Nanostructured in vitro TB latency model

纳米结构体外结核潜伏期模型

• 批准号: 7876788
• 负责人: GRAHAM S TIMMINS
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-19 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7876788
• 关键词: Aerobic; Animal Model; Anti-Infective Agents; Area; BCG Live; Bacillus (bacterium); Bacteria; Bacterial Model; Cells; Characteristics; Chemistry; Communities; Complement; Complex; Data; Disease; Drug Delivery Systems; Drug resistance; Figs - dietary; Film; Genus Mycobacterium; Goals; Growth; Human; Hybrids; In Vitro; Infection; Knowledge; Life; Lipids; Mediating; Metabolism; Methods; Modeling; Mus; Mycobacterium tuberculosis; Nanostructures; Nanotechnology; Oxygen; Pharmaceutical Preparations; Phenotype; Physiological; Preclinical Drug Evaluation; Publications; Recovery; Research; Resistance; Science; Silicon Dioxide; Solutions; Source; Structure; Testing; Time; Tuberculosis; Tuberculosis Vaccines; Work; X ray spectroscopy; X-Ray Diffraction; drug development; frontier; high throughput screening; implantation; in vitro Model; in vivo; in vivo Model; multidisciplinary; mycobacterial; nanobiology; nanostructured; novel; prevent; public health relevance; tool

DESCRIPTION (provided by applicant): Mycobacterium tuberculosis latently persist in 2 billion people, representing a major reservoir of tuberculosis (TB) due to reactivation, yet the state of latent bacilli remains a mystery. In order to eradicate TB, we need to understand latency and develop in vitro models for drug screening. In this interdisciplinary project we have merged microbiological tuberculosis research with nanotechnology and novel materials. Recent publications demonstrate a unique phenomenon of cell-directed assembly of hybrid lipid-inorganic nanostructures that drive a unique non-replicative persistence, and protect bacteria and drastically prolong viability. We show these same structures can be formed using mycobacteria, and that they display the highly extended viability characteristic of latency. We hypothesize that the persistence of latent TB, and its resistance to chemotherapeutic challenge, can be modeled by incorporation of bacilli into self-assembled lipid-inorganic nanostructures. This model combines the ability for in vitro study and growth condition modulation, with the potential for in vivo implantation and infection. We will generate a new in vitro model for high throughput screening for targets of, and drugs that can eliminate latent TB, to complement the existing complex in vivo models and al- low this field to move forward. Specific Aim 1 Define the conditions for optimal lipid-inorganic nanostructure-encasement of mycobacteria and its effects upon mycobacterial resistance to drugs. Specific Aim 2 Determine the changes in mycobacterial metabolism occurring upon encasement in lipid- inorganic nanostructures that result from non-replicative persistence, and drive in vitro survival and latency. Specific Aim 3 Demonstrate that lipid-inorganic nanostructure-encased mycobacteria represent valid models of latency by showing their infectivity in mice, even after extended periods of time after assembly. PUBLIC HEALTH RELEVANCE: Public Health Relevance We need to understand the way that tuberculosis is able to exist in a latent state because it is present in 2 billion people worldwide and reactivates into an active form in a significant percentage of cases. Clearing this 'reservoir' will be an essential component of an eventual eradication strategy. Although there are animal models of latency, there is no good model of in vitro latency, yet we need to have such an in vitro model so that we can understand how to attack it, and also to screen drugs with. We have developed a unique nanostructured model that mimics the extreme non-replicative persistence of latency, and want to test how effective it might be as a tool in the battle against TB.

描述（由申请人提供）：结核分枝杆菌潜伏存在于20亿人中，由于再活化，代表了结核病（TB）的主要储存库，但潜伏杆菌的状态仍然是一个谜。为了根除结核病，我们需要了解潜伏期并开发用于药物筛选的体外模型。在这个跨学科的项目中，我们将微生物结核病研究与纳米技术和新材料相结合。最近的出版物证明了混合脂质-无机纳米结构的细胞定向组装的独特现象，其驱动独特的非复制性持久性，并保护细菌并显著延长生存力。我们发现这些相同的结构可以使用分枝杆菌形成，并且它们显示出潜伏期的高度扩展的生存能力特征。我们假设潜伏性TB的持久性及其对化疗挑战的抗性可以通过将杆菌并入自组装脂质无机纳米结构来建模。该模型结合了体外研究和生长条件调节的能力，以及体内植入和感染的潜力。我们将建立一种新的体外模型，用于高通量筛选可以消除潜伏性TB的靶标和药物，以补充现有的复杂体内模型，并使该领域向前发展。具体目标1确定最佳脂质-无机纳米结构包裹分枝杆菌的条件及其对分枝杆菌耐药性的影响。具体目标2确定包裹在脂质无机纳米结构中时因非复制持续性而发生的分支杆菌代谢变化，并驱动体外存活和潜伏期。具体目标3证明脂质无机纳米结构包裹的分枝杆菌通过显示其在小鼠中的感染性代表有效的潜伏期模型，即使在组装后延长的时间段之后。 公共卫生相关性：我们需要了解结核病能够以潜伏状态存在的方式，因为它存在于全世界20亿人中，并在很大比例的病例中重新激活为活动形式。清除这一“水库”将是最终根除战略的一个重要组成部分。虽然有潜伏期的动物模型，但没有很好的体外潜伏期模型，但我们需要有这样一个体外模型，以便我们能够了解如何攻击它，并筛选药物。我们开发了一种独特的纳米结构模型，它模仿了极端的非复制性延迟持久性，并希望测试它作为对抗结核病的工具的有效性。